抗体抗原实验

The bimolecular fluorescence complementation (BiFC) allows not only the investigation of protein interactions but also the visualization of protein complexes in living cells. This method is based on two nonfluorescent fragments of fluorescent proteins (FPs) which can reassem ...

Co-affinity purification methods can test whether two proteins physically engage in a complex. The assay principle is to enrich cellular extracts for a first protein by a purification step, and then test if a second protein is enriched as well. This principle has been optimized for use at high-thr ...

The production of cytokines is a crucial element of the host response to viral and bacterial infections. To follow these events in vivo, transgenic mice have become a valuable tool to study cytokine production through induction of reporter genes. We describe here the generation and immortal ...

The completion of human and mouse genome sequencing has confronted us with huge amount of data sequences that certainly need decades and many generations of scientists to be reasonably interpreted and assigned to physiological functions, and subsequently fruitfully translated i ...

The ability to introduce DNA sequences (e.g., genes) of interest into the germline genome has rendered the mouse a powerful and indispensable experimental model in fundamental and medical research. The DNA sequences can be integrated into the genome randomly or into a specific locus by homol ...

Previous studies have demonstrated that dendritic cells (DC), macrophages (Mφ), and their precursors monocytes are susceptible to infection by human cytomegalovirus (HCMV) in the natural host as well as in vitro. Due to their proficient ability to take up and present antigens to the lymphoc ...

Natural killer (NK) cells are major components of the innate immune system and are assumed to play an important role in the defense against cytomegalovirus infection. Human cytomegalovirus (HCMV) is the only virus known that shapes the NK cell receptor repertoire in humans. Assays allowing t ...

Natural Killer (NK) cells are crucial to the control of many viral infections. They are able to kill infected cells directly through the secretion of cytotoxic granules or through binding to death receptors on target cells. They also secrete cytokines and chemokines and, through interactio ...

The yeast two-hybrid (Y2H) system is a powerful method to identify and analyze binary protein interactions. In the field of virology, the Y2H system has significantly increased our knowledge of structure and function of viral proteins by systematically assessing intraviral protein in ...

CMV-specific T cells may be detected and quantified after antigen-specific stimulation based on the induction of cytokines as a readout system. Secreted cytokines may be detected from the supernatant of stimulated cells using ELISA. Alternatively, antigen-specific cytokine-s ...

Herpes simplex encephalitis (HSE) is one of the most severe viral infections affecting the temporal lobes of the brain. Despite the improvements in diagnosis and antiviral drug treatment, one third of all patients fail to respond to therapy or subsequently suffer neurological relapse and ...

Infection with herpes simplex virus type 1 (HSV-1) causes the most common skin disease. Various test systems have been established to recapitulate this cyclical pathway of productive infection, latency, and reactivation. Most studies of latency and reactivation are conducted in anim ...

In this paper we show how to obtain a three-dimensional model of virus-infected cells by serial sectioning of resin embedded samples and transmission electron microscopic imaging. The method bases on sample fixation by high pressure freezing and processing by freeze substitution with ...

Imaging host–pathogen interactions in real time can provide significant insight into dynamic processes and provide information about time and space of their occurences. Here, we present detailed experimental instructions on how to image the membrane penetration process of the non ...

The existence of cell type-specific entry pathways of human cytomegalovirus is an unresolved question as the course of viral entry in different cell types is still not fully understood. To gain more insight into these processes, we generated a dual fluorescent HCMV, where the capsid-associ ...

miRNAs have in recent years emerged as novel players in virus–host interactions. While individual miRNAs are capable of regulating many targets simultaneously, not much is known about the role of distinct host or viral miRNAs in the context of infection. Analysis of the function of a miRNA is often ...

Quantification of infectious virus is crucial to many experimental approaches in virological research. A broadly used and facile technique is the so-called “plaque assay” which provides precise information on the absolute quantity of infectivity in a given volume. Due to advances in the ...

Virology has greatly benefited from the introduction of fluorescent proteins (FP’s) as tags to viral as well as cellular structures. With advanced imaging technologies it is now possible to observe host–pathogen interactions in living cell systems in real-time. The generation of high- ...

Fluorescence in situ hybridization (FISH) is widely used to visualize nucleotide sequences in interphase cells or on metaphase chromosomes using specific probes that are complementary to the respective targets. Besides its broad application in cytogenetics and cancer researc ...

Multimeric antibody fragments, particularly dimers (diabodies), trimers (triabodies), and tetramers (tetrabodies) of single-chain Fv molecules (scFv), provide high avidity through multivalent binding to the target antigen. The combination of their smaller size and avid bin ...